Structure, organization, and transcription of Drosophila U6 small nuclear RNA genes

Abstract

U6 RNA is an abundant, capped small nuclear RNA (snRNA) associated with hnRNP particles (Reddy, R., and Busch, H. (1983) Prog. Nucleic Acid Res. Mol. Biol. 30, 127-162). Small nuclear ribonucleoprotein particles containing U4 and U6 RNAs are required components for splicing of pre-mRNAs (Berget and Robberson, 1986; Black and Steitz, 1986). In this study the Drosophila U6 RNA genes have been isolated and characterized. The Drosophila genome contains three U6 snRNA genes which are clustered in a 2-kilobase-pairs long DNA fragment. The U6 RNA coding regions are 100% homologous in all three genes, but the flanking sequences diverged significantly from each other. A possible secondary structure model for the Drosophila U4/U6 RNA complex is presented. Consistent with our previous observation that U6 RNA is a RNA polymerase III product (Reddy, R., Henning, D., Das, G., Harless, M., and Wright, D. (1987) J. Biol. Chem. 262, 75-81), all three genes contained a region homologous to the consensus intragenic regulatory region and a cluster of T residues on the 3'-end, characteristic of genes transcribed by RNA polymerase III. A TATA box was found between nucleotides -23 and -31, and a stretch of 28 nucleotides from -43 to -71 was conserved in the 5'-flanking region of all three U6 RNA genes. The Drosophila U6 RNA genes were transcribed in vitro by Drosophila nuclear extracts but were not transcribed by Novikoff hepatoma or HeLa cell extracts. Similarly, a mouse U6 RNA gene was transcribed in Novikoff hepatoma or HeLa cell extracts but not in Drosophila nuclear extracts. These results suggest that species-specific factor(s) are involved in the transcription of U6 snRNA genes.